20020012002

201201120122013

RPF-I

PROFORMA FOR SUBMISSION OF RESEARCH PROJECTS

PART -I: GENERAL INFORMATION

Project CodeInstitute code No.ICAR Code No.

Name of the Institute and DivisionName and address of InstituteName of Division/ SectionLocation of project

CI

IGFRI, JHANSI-284 003Crop Improvement DivisionJhansi & Avikanagar

202 Project TitleDevelopment of drought-tolerant stay-green forage pearl millet

2032031

2042041

20520512052

2062061

Priority AreaResearchapproach

Applied ResOJ.

Genetic ImprovementBasic Res Process/Technology

02 Development03

Transfer ofTechnology

04

Specific AreaPrevious project/projects in thisspecific area (Year, type of funding,cost etc.)

DurationDate of startLikely date of completion

Total cost of the projectForeign exchange component (if any)

Forage crop improvement

5 years.,2.O.l-G-H- :20 II - J 2-2015-16

Nil

207 Project profile summaryPearl millet [Pennisetum glaucum (L.) R. Br.] is a cross-pollinating C4 cerealcrop of the hottest, driest areas of the tropics and subtropics with the highestwater-use efficiency. It can provide grazing, green fodder in varying cuttingregimes or silage, while its stover is a valuable source of feed, making it a dual-purpose crop for subsistence farmers. The stover, however, remains green tothe point of grain harvest, thus lifting its nutritional value above that of othercereal residues, such as maize stover. Pearl millet fodder when fed green to theanimals has easy digestibility than the chaffed straw. It produces grain andfodder under very hot and dry conditions, and on soils too poor for sorghumand maize. It is usually grown as a dryland dual-purpose grain & fodder crop insemi arid regions of Africa & India, although it is sometimes irrigated in India.P.~~r.I..mjJJ.~~-'..\:l.Q~~.Y.~x,.r.~~m~mg~.tQ.fc:.t.Y.9.I,lX~R!~.(~r~j!j~y..c:.t.D.cJ..mQj~t!-!r.~CfQJ}9.i.~iQ'1~.

Agricultural drought may develop at any time of the cropping season. Droughtavoidance 'stay-green' trait in sorghum and drought tolerance 'high paniclethreshing percentage' trait in pearl millet have been found to be associated withpost-flowering [terminal] drought tolerance. Th~.g9.c:.tJ..QUhj~..assignedresearchpr9j~~.~"Development of drought-tolerant stay-green forage pearl millet" isro..m~t\~..p.hy.~.iQ!Qg.i.G~J..t.mjt.~!~y':-g~~~.I)..y.~!!.mR!~.f-Q~.gm.~n.~m!.(9.9.Q~X.prQQ!-!~tjQD!-m5:J.~r.... .w.c:.t.~~r:-.I.i.mj!~g.....~~mgjti.Qn~. Through Institute multi-disciplinarycollaborations, research will target genetic traits (such as phenotypic,anatomical, physiological & biochemical) of pearl millet for improved forageyield and quality.

208 Key words: Forage pearl millet, drought tolerance, stay-green.

210 Principal Investigator,2101 Name2102 Designation2103 Division/Section2104 Location2105 Institute Address

211 Co-investigator2111Name2112 Designation2113 Divlsion/Section2114 Location2115 Institute Address

PART-II: INVESTIGATOR PROFILE

A.K. SinghSr. Scientist, Plant BreedingCrop Improvement DivisionJhansiIndian Grassland and Fodder Research InstituteJhansi-284 003, Uttar Pradesh

S.S. MeenaPrincipal Scientist, Plant BreedingCrop ImprovementAvikanagarIndian Grassland and Fodder Research InstituteRegional StationAvikanagar-304 501, Rajasthan

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F

2202201

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PART - III : TECHNICAL DETAILS

Introduction and objectives:Origin of the project: (Problem identification)In the crops of the semi-arid tropics the stay-green character is of vitalimportance in the struggle to feed human and livestock. Disease and insectresistance, wide environmental adaptability, appropriate time to maturity,good tillering, and reasonably good grain yield as well as juiciness,palatability and digestibility of crop residues are the major criteria ofimproving the sorghum and millet crops of the semi arid zone. Pearl milletis a dual purpose crop, being important for its grain in Africa and Asia. Itis considered as one of the most drought-tolerant domesticated cereals.India is the largest producer of this crop and its productivity andproduction have increased, despite declining area. Forage pearl millets areharvested principally by pasturing, green chopping Among abiotic andbiotic stresses, drought and downy mildew where identified as mostimportant in that order. Sorghum and millet lines exhibiting the stay-greenphenotype often show enhancement of other desirable characters as well.Stay-green is a mechanism of drought tolerance characterized by theretention of green leaf area at drop maturation under drought stress, but theexpression of stay-green properties varies among plant species.

2202 Definition of problem:A single mechanism cannot accord complete drought resistance to agenotype. Almost all phases of forage pearl millet life cycle, fromgermination through vegetative and reproductive development are subjectto drought stress in the arid and semi-arid regions of South Asia andSahelian-Sudanian zone of Africa receiving an annual rainfall of 200-800mm and are influenced by programmed cell death.

2203 Immediate objectives:1. To identify stay-green trait associated with drought tolerance in Pearl

millet.2. To develop drought-tolerant stay-green forage Pearl millet population

for terminal drought tolerance.

2204 Long term objectives:1. To introgress the stay-green trait into superior pearl millet cultivars for

further improving drought tolerance and nitrogen use efficiency.

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2205 Review of status of research in the subject:a) International StatusP.m~gQ.t~.tf~~~can occur any time during the crop cycle, severe moisturestress during emergence and the early seedling phase causes seedling deathand poor crop establishment. Poor or uneven crop stands are some of themajor causes for low biological yield and high pearl millet yield losses(Soman et al. 1987). Stress occurring after crop establishment but withinthe seedling phase has little effect on plant survival or grain yield (Lahiriand Kharbanda 1965; Lahiri and Kumar 1966). Mahalakshmi and Bidinger(1985, 1986) concluded that water stress during vegetative phase reducedto the dominance of the main shoot and allowed additional tillers tocomplete their development. Mahalakshmi et al (1987) further establishedthat stress during the vegetative growth phase delayed flowering of themain shoot, which increased the chances for escape from water stress.Lahiri and Kharbanda 1965; Lahiri and Kumar 1966; Mahalakshmi andBidinger (1985); Mahalakshmi et at. 1987 found that pearl millet was mostsensitive to water stress during flowering and grain filling stages and therewas yield reduction due to decrease in no. of panicles per plant and adecrease in grain mass; the latter appeared to be mainly due to ashortening of the grain filling period rather than to a reduction in graingrowth rate (Bieler et al. 1983). Pearl millet has the capacity tocompensate for such a reduction by mobilizing stored soluble sugars(Fussell et al. 1980). Drought tolerant Pearl millet varieties HB 5, K 559showed higher germination than the susceptible varieties Tift 23 DA x Tift18 DB and Tift 23 DA x Tift 383 at a higher osmotic pressure (Saint Clair1980).

Leaves are genetically programmed to die and their senescence can beinitiated by environmental cues as part of an adaptive strategy. Theclearest symptom of ~~D~~9.Y.l).9~in leaves is the visible loss of green colourwith time. Chloroplasts become dispensable during senescence. Thechlorophyll degradation pathway is turned on, stored proteins are brokendown and photosynthesis declines, reflecting the physiological decline.Delayed senescence or stay-green (Borrell et at 2003; Mahalakshmi &Bidinger 2002; Thomas & Howarth 2000) is a mechanism of droughttolerance characterized by the retention of green leaf area at crop maturityunder water-stressed environments (Borrell et al 2000). The stay-greentrait in sorghum was found genetically and physiologically complex,expressing a variety of patterns and environmental sensitivities dependingon the background genotype (Thomas & Howarth 2000).

St~y':-gf~~1]is a mechanism of drought tolerance characterized by theretention of green leaf area at drop maturation under drought stress, but theexpression of stay-green properties varies among plant species. Forexample, the record yield of maize grain, almost 240 q/ha, was achieved in1985 on a farm of Illinois with the variety FS854, a stay-green maize line.Stay-green is the best-characterized component of drought toleranceavailable in sorghum (Mahalakshmi and Bidinger 2002). Green leaf areaafter flowering has been related to grain yield under moisture stress inwheat (Fischer and Kohn 1966) and maize (Tollenaar and Daynard 1978).Stay-green therefore has been suggested as an indirect selection criterionfor post-flowering drought tolerance (Rosenow et at. 1983).

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Knowledge about the inheritance of stay-green is a prerequisite for the successfuluse of this trait as a selection criterion. Non-senescence in sorghum, measured asgreen leaf area (GLA) retention, was reported to be regulated by both dominantand recessive epistatic interactions (Tenkouano et al. 1993). The level of dominantgene action, however, depends on the environment (Walulu et al. 1994). Keepingleaves alive for longer is a fundamental strategy for increasing crop production,particularly under water-limited conditions.

Scientific literature concerning stay-green in Pearl millet is very less compared toSorghum. In pearl millet, the expression of relative chlorophyll content in theFiof a putative stay-green inbred 02F 266-4 and a normal senescent line Tift 454indicated stay-green a dominant or over-dominant trait. The QTL mappingapproach is also revealing details of the genetic and environmental control of leafsenescence in pearl millet. Howarth et al. 1994 and Yadav et al. 1999 produced across between non-senescent line (H 77 833-2) and senescent line (PRL T 2/89-33)of pearl millet in order to develop a mapping family to study drought-relatedsenescence and other traits further. It is currently being investigated whether pearlmillet resembles sorghum in an association between stay-green and maintenanceof grain yield under post-flowering drought stress. .

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2205 Review of status of research in the subject:b) National StatusSorghum and Pearl millet crops currently constitute an estimated 11.4% ofthe global cereal area harvested and 4.1 % of total output of world cereals(FAa 2009). Kellog et al (1996) observed that farmers in India rejectedseveral Pearl millet cultivars improved only for grain yield because of toopoor fodder value of the stover. The soil crust formation due to rainfallimmediately after sowing is a serious problem to seedling emergence(Sharma & Agarwal 1979). Yadav and am Prakash (2001, 2003)developed a simple and low cost workable rainout shelter for croppingsystems research under rainfed farming. Joshi et al. (2005) evaluated pearlmillet hybrids for their tolerance to high temperature and limiting soilmoisture at the seedling stage and their results indicated that seedlinggrowth parameters such as shoot dry mass and leaf elongation rate and thechlorophyll stability index (CSI) can be used for screening against hightemperature and limited soil moisture stresses by stopping irrigation afterboot leaf stage until maturity and found significant positive corelateions ofyield with days to flowering and drought susceptibility index. Babu et al.(1995), am Prakash et al. (2008) studied the effect of prolong droughts of15 days at different growth stages of pearl millet and found tillering stagethe most susceptible to drought, followed by drought at grain filling andmaturity stage. Upadhyay and Murty (1971) observed that pearl milletgenetic stocks which were good for vegetative characters were deficient inreproductive traits and vice-versa. Gupta et al. (1975) found that there wasoccurance of seasonal and genotype x environmental interactions forfodder yield and sum of its components. Weltzien et a.l (1996) found indiscussion that farmers in western Rajasthan perceived drought as themain cause for Pearl millet crop failures and yield losses; however, thereappeared to be a strong interaction with soil fertility. Gene effects wereprimarily additive for most of the green fodder traits in Pearl millet (Gupta1995). According to Joshi et al (1999) terminal drought stress in Pearlmillet is found to be the most detrimental factor to the growth anddevelopment of grain and ultimately results in considerable loss in grainyield. Tiwana et al. (2003) and Suneetha Devi and Padmaja 2007 reportedincrease in forage yield of Pearl millet with increased levels of nitrogenand its favorable effects on growth parameters. Govindaraj et al. (2011)studied genetic diversity in indigenous Pearl millet accessions based onbiometrical and nutritional quality traits and found that presence ofsignificant genetic variability can be exploited through hybridization ofgenotypes from different clusters and subsequent selection from thesegregating generations. Yadav et al (2002) studied quantitative trait lociassociated with traits determining grain and stover yield in Pearl milletunder terminal drought-stress conditions. Bidinger et al (1987) suggestedearliness as a mechanism of escape from drought in Pearl millet; selectionfor grain yield under terminal moisture stress can help in selection forcombination of high yield and adaptation to terminal drought. Accordingto Bidinger et al (1987), there is an evidence for genetic differences inadaptation to drought stress during emergence and establishment in Pearlmillet; direct selection for these traits is possible and must be givenemphasis while breeding for drought resistance as an objective.

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221 Project Technical Profile:2211 Organization of work elements(For each objectives and participating investigator giving man-months involved)----------.-----.-._-Investigator Designation Contributions Man-monthsBreeder Sr. Scientist, Planning, acquisition and assessment 7(A.K. Singh) PI. Breeding of genetic material, screening for

stay-green, conducting conventionalbreeding for drought-tolerant stay-green forage pearl millet.

Breeder (allotted) Pri. Scientist, Acquisition and assessment of 4(Dr. S.S. Meena) PI. Breeding genetic material, and evaluation.

Physiologist Developing simple and reliable 3(in-principle) screening technique for drought

tolerance using stay-green trait,estimating chlorophyll content,green leaf area retention, leaf area,leaf temperature and studyingchlorophyll metabolism.

Soil Scientist! Measuring soil moisture status and 3Agronomist soil temperature during different(not allotted) phenological stages under drought

and normal condition.

2212 Methodology:A)

1) Standardization of screening technique using stay-green trait.2) Identification of populations for stay-green trait under post-flowering drought in

post-rainy season.3) Study of inheritance of stay-green in pearl millet.4) Hybridizing putative stay-green pearl millet with superior cultivars and selection.5) Comparison of chlorophyll content using chlorophyll meter and physiological

analysis.6) Evaluation of selected population for drought tolerance and stay-green.

2213 Plan of actionIt depends based on availability of project resources.

2214 Time schedule of activities (milestone)i) Acquisition and assessment of germplasm. [From 1st yr. onward)'ii) Comparison of chlorophyll content using chlorophyll meter and physiological

analysis. [1st yr. onward]iii) Standardization of screening technique using stay-green trait. [2nd yr. & 3rd yr.]iv) Identification of populations for stay-green trait under drought in post-rainy

season. [2nd yr. & 3rd yr.]v) Hybridizing putative stay-green pearl millet with superior cultivars and selection.

[2nd yr. onward]vi) Evaluation of selected population for drought tolerance and stay-green. [5th yr.]

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First yeari)ii)

2215 Annual targets for each activity:

Acquisition and assessment of germplasm.Comparison of chlorophyll content using chlorophyll meter and physiologicalanalysis.Preliminary selection for stay-green in pot culture.iii)

Second Yeari) Acquisition and assessment of germplasm.ii) Comparison of chlorophyll content using chlorophyll meter and physiological

analysis.iii) Standardization of screening technique using stay-green trait.iv) Identification of populations for stay-green trait under drought in post-rainy

season.v) Hybridizing putative stay-green pearl millet with superior cultivars and

selection. .

Third Yeari) Acquisition and assessment of germplasm.ii) Hybridizing putative stay-green pearl millet with superior cultivars and

selection.iii) Identification of populations for stay-green trait under drought in post-rainy

season.

Fourth Yeari) Hybridizing putative stay-green pearl millet with superior cultivars and selection.

Fifth Yeari) Hybridizing putative stay-green pearl millet with superior cuItivars and selection.ii) Evaluation of selected population for drought tolerance and stay-green.iii) Project report preparation and submission.

2216 Estimated man monthsa) Scientific: 17b) Technical: 12c) Supporting: 12 (

222 Proposed Research Details2221 Importance of the project (gaps in knowledge/ products/ process technology) to the

institute mandate.

First Year [2011]1. Acquisition & assessment of the requisite genetic material (stay-green & tillering,landraces, inbreds, varieties), and multiplication for seed increase.2. Preliminary phenotypic selection for delayed leaf senescence in pot culture undermoisture stress.

Second Year3. Identification of populations for stay-green trait under drought during post-flowering for terminal drought tolerance --- by conducting field experiments at two

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locations (Jhansi & Avikanagar) with two water regimes after flowering in the post-rainy season

First water regime (moisture stress) by withholding irrigationSecond water regime (free from moisture stress) by irrigating the crop till

physiological maturityCorrelation between green leaf retention during different post-flowering stages withbiomass and grain yield.4. Comparison of chlorophyll content of a putative stay-green and non-stay-greenpopulations over time.

Third Year5. [Before Kharifseason] Hybridization between improved forage varieties andpopulation with stay-green trait at both locations.6. [In Kharif season] Single-plant screens in the F2 generation in large number ofpopulations for stay-green trait under drought for terminal drought tolerance at bothlocations.7. Comparison of chlorophyll content in screened single plants for identification ofstay-green traits.

Fourth Year8. [Before Kharif season] Single-plant screens in the F3 generation for stay-greentrait under drought for terminal drought tolerance at both locations.9. Comparison of chlorophyll content in screened single plants for identification ofstay-green traits.10. [In Kharifseason] Making screens in the F4 generation for identification ofstay-green populations associated with drought tolerance at both locations.

Fifth Year11. [Before Kharif season] Making screens in the F5 generation for identification ofstay-green populations associated with drought tolerance at both locations.12. Comparison of chlorophyll content in screened single plants for identification ofstay-green traits.13. [In Kharif season] Evaluation of populations exposed to drought foridentification of drought-tolerant stay-green forage Pearl millet at both locations.

Proposed ParametersGrowth parameters [plant height, days to 50% flowering, days to complete earing,days to maturity, biomass]

Physiological parameters [chlorophyll content, green leaf area retention, leaf areaper plant, No. ofleaves per plant, leaf temperature, photosynthetic capacity]Morphological and Forage parameters [stem thickness, leafiness, No. of tillers, plantheight, Leaf/stem ratio, less seed production potential, nutrient quality]

2222 stions attempted to be answered:i) Is it possible to develop drought-tolerant stay-green pearl millet for forage in

within a half decade?

2223 Anticipated Process/ Products/ technology/ Knowledge Expected to be evolvedby pursuing the project:If developed the stay-green trait should have multiple benefits in dual-purposepearl millet improvement.

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2224 Practical Utility of anticipated results of the project:a) Immediate benefits.

Standardization of drought tolerance screening technique using stay-green for useby researchers.

b) Medium Term benefits.Identification of putative stay-green trait in pearl millet and no extra cost incultivation of stay-green forage pearl millet.

c) Long Term Benefits.Better understanding of senescence in pearl millet.

2225 Expertise available with investigator group/ Institute.Scientists available in the Institute IGFRI.

2226 Expertise (if any) to be obtained by investigator group from outside theInstitute.

a. Within Countryb. Outside Country

Not possibleNot possible

PART- IV: BUDGET ESTIMATES(Summary)

230 Budget Summary(Recurring)

\, /

2301 Salaries Year (1) Year (2) Year (3) TotalScientificTechnicalSupportingWagesSub Total

2302 Consumables:I. Chern icalsii. Glasswaresiii. Others

Sub Total2303 Travel2304 Miscellaneous2305 Sub Total (Recurring)231 Non-Recurring Expenditure

( Equipments)232 Total (230&231)233 Salaries / Wages:2331 Justification (for technical/wages/labour in terms of work content)234 Consumables Rs. 2 lakhs2341 Justification235 Travel Rs. 50,0002351 Justification236 Other Cost/ Miscellaneous Year 1 Year2 Year 32361 Field preparation/ planting/ harvesting (man days/ cost)2362 Inter cultivation (man days/ cost)2363 Animal maintenance2364 Any other items2365 Justification for above237 Equipment

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2371 Equipment already available to be used in project2372 Equipment to be purchased with cost: ---2373 Justification for each additional equipment2374 Equipment to be imported2375 Justification for import238 Additional infrastructure facilities (if needed)2381 Works2382 Land (sq. meter)2383 Animals239 Financing OrganizationIf funded by an organization other than the institute, then give the following information:

(a) Name the financing organization:(b) Title of the project

(if the project forms a part ofa larger project)

PART-V: DECLARATIONThis is to certify that• The research work proposed in the schemel project does not in any way duplicate the

work already done or being carried out in the institute on the subject.• The same project has been/ has not been submitted to any other agency(ies) for

financial support(if already submitted identify project and agency).• The Investigator/co-investigators have been fully consulted in the development of

project and have fully undertaken the responsibility to carry out the programme as perthe technical progra e.

~ ~,~(S.S. Meena)

Co Project Investigator

(A.K.Si~ blPrincipal Investigator

~ Q) ~lL.J, ~ .. ~. f~~~. 0a Signature and comments oftbe Head of the Division/ Section ~t::...-----~-~

"2..:!> I g1 ''L---Signature and comments of the Joint Director (Research)

Signature and comments of the Director

II